Willifordwade6598

Post-translational histone modifications and chromatin remodelling play a critical role controlling the integrity of the genome. Here, we identify histone lysine demethylase PHF2 as a novel regulator of the DNA damage response by regulating DNA damage-induced focus formation of 53BP1 and BRCA1, critical factors in the pathway choice for DNA double strand break repair. PHF2 knockdown leads to impaired BRCA1 focus formation and delays the resolution of 53BP1 foci. Moreover, irradiation-induced RPA phosphorylation and focus formation, as well as localization of CtIP, required for DNA end resection, to sites of DNA lesions are affected by depletion of PHF2. These results are indicative of a defective resection of double strand breaks and thereby an impaired homologous recombination upon PHF2 depletion. In accordance with these data, Rad51 focus formation and homology-directed double strand break repair is inhibited in cells depleted for PHF2. Importantly, we demonstrate that PHF2 knockdown decreases CtIP and BRCA1 protein and mRNA levels, an effect that is dependent on the demethylase activity of PHF2. Furthermore, PHF2-depleted cells display genome instability and are mildly sensitive to the inhibition of PARP. Together these results demonstrate that PHF2 promotes DNA repair by homologous recombination by controlling CtIP-dependent resection of double strand breaks. © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.The spatiotemporal regulation of chromosome segregation and cell division in Caulobacter crescentus is mediated by two different P-loop ATPases, ParA and MipZ. Both of these proteins form dynamic concentration gradients that control the positioning of regulatory targets within the cell. Their proper localization depends on their nucleotide-dependent cycling between a monomeric and a dimeric state and on the ability of the dimeric species to associate with the nucleoid. In this study, we use a combination of genetic screening, biochemical analysis and hydrogen/deuterium exchange mass spectrometry to comprehensively map the residues mediating the interactions of MipZ and ParA with DNA. We show that MipZ has non-specific DNA-binding activity that relies on an array of positively charged and hydrophobic residues lining both sides of the dimer interface. Extending our analysis to ParA, we find that the MipZ and ParA DNA-binding sites differ markedly in composition, although their relative positions on the dimer surface and their mode of DNA binding are conserved. In line with previous experimental work, bioinformatic analysis suggests that the same principles may apply to other members of the P-loop ATPase family. P-loop ATPases thus share common mechanistic features, although their functions have diverged considerably during the course of evolution. © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.Previously, we have shown that human sperm Prohibitin (PHB) expression is significantly negatively correlated with mitochondrial ROS levels but positively correlated with mitochondrial membrane potential and motility. However, the possible role of PHB in mammalian spermatogenesis has not been investigated. Here we document the presence of PHB in spermatocytes and its functional roles in meiosis by generating the first male germ cell-specific Phb-cKO mouse. Loss of PHB in spermatocytes resulted in complete male infertility, associated with not only meiotic pachytene arrest with accompanying apoptosis, but also apoptosis resulting from mitochondrial morphology and function impairment. Our mechanistic studies show that PHB in spermatocytes regulates the expression of STAG3, a key component of the meiotic cohesin complex, via a non-canonical JAK/STAT pathway, and consequently promotes meiotic DSB repair and homologous recombination. Selleckchem SM04690 Furthermore, the PHB/JAK2 axis was found as a novel mechanism in the maintenance of stabilization of meiotic STAG3 cohesin complex and the modulation of heterochromatin formation in spermatocytes during meiosis. The observed JAK2-mediated epigenetic changes in histone modifications, reflected in a reduction of histone 3 tyrosine 41 phosphorylation (H3Y41ph) and a retention of H3K9me3 at the Stag3 locus, could be responsible for Stag3 dysregulation in spermatocytes with the loss of PHB. © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.Hand burns are common and often complex injuries, requiring referral to specialist centers. The patient's thumbprint is a rapid means of accurately assessing hand burn surface area. This study aimed to establish categories and evaluate sites of hand burn surface area in order to facilitate comparison of hand burns. Sixteen burns involving the hand and wrist among 14 patients referred to a burns center were retrospectively categorized by burn mechanism, burn thickness, and hand burn surface area. The burn surface area in the clinical record was compared with that calculated by the thumbprint method and the rule of thumb diagram. Burn surface area in the clinical record was either "1%" or " less then 1%" in 9 of 16 cases. In contrast, the surface area was less than one thumbprint (T) in 6 burns, two were between 1 and 5T, three were 5-10T, two were 10-20T, two were 20-50T and one was greater than 50T. Median thumbprint burn surface area was 1.5T (range 0.20-80T), which corresponds to 0.05% TBSA. The hand areas with the highest burn frequency per unit area were the dorsum of the hand and dorsum of the index finger, with relative sparing of the palm and palmar surface of the digits. Hand burns surface area varies widely, and thumbprint evaluation with categories and mapping allows finer distinction between the surface area proportions and specific sites involved, even in a small series of hand burns. © The Author(s) 2020. Published by Oxford University Press on behalf of the American Burn Association. All rights reserved. For permissions, please e-mail journals.permissions@oup.com.PURPOSE To identify potential performance indicators relevant for district healthcare systems of Ethiopia. DATA SOURCES Public Library of Medicine and Agency for Healthcare Research and Quality of the United States of America, Organization for Economic Cooperation and Development Library and Google Scholar were searched. STUDY SELECTION Expert opinions, policy documents, literature reviews, process evaluations and observational studies published between 1990 and 2015 were considered for inclusion. Participants were national- and local-healthcare systems. The phenomenon of interest was the performance of healthcare systems. The Joanna Briggs Institute tools were adapted and used for critical appraisal of records. DATA EXTRACTION Indicators of performance were extracted from included records and summarized in a narrative form. Then, experts rated the relevance of the indicators. Relevance of an indicator is its agreement with priority health objectives at the national and district level in Ethiopia. RESULTS OF DATA SYNTHESIS A total of 11 206 titles were identified. Finally, 22 full text records were qualitatively synthesized. Experts rated 39 out of 152 (25.7%) performance indicators identified from the literature to be relevant for district healthcare systems in Ethiopia. For example, access to primary healthcare, tuberculosis (TB) treatment rate and infant mortality rate were found to be relevant. CONCLUSION Decision-makers in Ethiopia and potentially in other low-income countries can use multiple relevant indicators to measure the performance of district healthcare systems. Further research is needed to test the validity of the indicators. © The Author(s) 2020. Published by Oxford University Press in association with the International Society for Quality in Health Care. All rights reserved. For permissions, please e-mail journals.permissions@oup.com.Cervical mucus produced by the endocervix plays an essential role as a hormonally induced regulator of female fertility. Cervical mucus fluctuates in both physical characteristics and in sperm penetrability in response to estrogens and progestogens. However, the mechanisms by which steroid hormones change mucus remains poorly understood. Current in vitro models have limited capability to study these questions as primary endocervical cells possess limited expansion potential, and immortalized cells lose in vivo characteristics such as steroid sensitivity. Here we overcome these limitations by establishing an in vitro primary endocervical cell culture model using conditionally reprogrammed cells (CRCs). CRC culture utilizes a Rho-kinase inhibitor and a fibroblast feeder layer to expand proliferative potential of epithelial cell types that have normally short in vitro life spans. In our studies, we produce CRC cultures using primary endocervical cells (CRECs) from adult female rhesus macaques (Macaca mullata). We demonstrate that primary endocervical cells from the nonhuman primate (NHP) can be robustly expanded using a CRC method, while retaining steroid receptor expression. Moreover, when removed from CRC conditions and switched to differentiation conditions, these cells are able to differentiate and produce mucus including MUC5B, the most prevalent mucin of the endocervix. We conclude that this method provides a promising in vitro platform for conducting mechanistic studies of cervical mucus regulation as well as for screening new therapeutic targets for fertility regulation and diseases of the endocervix. © The Author(s) 2020. Published by Oxford University Press on behalf of Society for the Study of Reproduction.OBJECTIVE To know what hospital managers and safety leaders in Ibero-American countries are doing to respond effectively to the occurrence of adverse events (AEs) with serious consequences for patients. DESIGN Cross-sectional international study. SETTING Public and private hospitals in Ibero-American countries (Argentina, Brazil, Chile, Colombia, Mexico, Peru, Portugal and Spain). PARTICIPANTS A convenience sample of hospital managers and safety leaders from eight Ibero-American countries. A minimum of 25 managers/leaders from each country were surveyed. INTERVENTIONS A selection of 37 actions for the effective management of AEs was explored. These were related to the safety culture, existence of a crisis plan, communication and transparency processes with the patients and their families, attention to second victims and institutional communication. MAIN OUTCOME MEASURE Degree of implementation of the actions studied. RESULTS A total of 190 managers/leaders from 126 (66.3%) public hospitals and 64 (33.7%) private hospitals participated. Reporting systems, in-depth analysis of incidents and non-punitive approaches were the most implemented interventions, while patient information and care for second victims after an AE were the least frequent interventions. CONCLUSIONS The majority of these hospitals have not protocolized how to act after an AE. For this reason, it is urgent to develop and apply a strategic action plan to respond to this imperative safety challenge. This is the first study to identify areas of work and future research questions in Ibero-American countries. © The Author(s) 2020. Published by Oxford University Press in association with the International Society for Quality in Health Care. All rights reserved. For permissions, please e-mail journals.permissions@oup.com.